Specific primers and liquid-phase chip for SNP (Single Nucleotide Polymorphism) detection of chromosome 4q25 section

Abstract

The invention discloses specific primers and a liquid-phase chip for SNP (Single Nucleotide Polymorphism) detection of a chromosome 4q25 section. The liquid-phase chip comprises an ASPE primer, anti-tag sequence coated microspheres, and an amplification primer, wherein the ASPF primer is composed of a 5'-terminal tag sequence and 3'-terminal specific primers for target gene mutation, and the specific primers are respectively SEQ ID NO. 11 and SEQ ID NO. 12 specific for a C172T SNP site, SEQ ID NO. 13 and SEQ ID NO. 14 specific for a C133A SNP site, SEQ ID NO. 15 and SEQ ID NO. 16 specific for a T139G SNP site, SEQ ID NO. 17 and SEQ ID NO. 18 specific for a T80A SNP site, and / or SEQ ID NO. 19 and SEQ ID NO. 20 specific for a T105C SNP site. According to the invention, the coincidence rate between the detection result of the liquid-phase chip for SNP detection of the chromosome 4q25 section and the detection result of a sequencing method is up to 100%, not only is the condition of single site mutation detected, but also the polymorphism conditions of multiple mutant sites can be simultaneously detected in parallel, and the detection effects are consistent.

Description

technical field [0001] The invention belongs to the field of molecular biology, relates to medicine and biotechnology, in particular to a specific primer for detecting SNP in the segment of chromosome 4q25 and a liquid phase chip. Background technique [0002] Band 5, region 2, long arm of human chromosome 4 (4q25) is a "gene desert" region with a length of about 15 million base pairs, and the closest functional gene to 4q25 is paired-like homeodomain transcription factor 2 (Paired- like homeodomain transcription factor 2, PITX2), which is also 5000bp away from 4q25. PITX2 is considered to be a key transcription factor regulating the left-right asymmetry of internal organs in vertebrates, and plays a key role in the development of cardiac embryos, affecting the configuration of the left atrium. [0003] The 4q25 sequence does not produce a protein, but it is not meaningless and may contain disease-related information or have other important roles in human physiology. Studi...

AI Technical Summary

Problems solved by technology

The PCR-RFLP method is based on the change of the restriction endonuclease recognition site caused by gene mutation, such as the loss of the site or the generation of a new site, a specific fragment is amplified by PCR, and then cut and amplified with a restriction endonuclease. The size of the fragment is observed by electrophoresis. This method is used to detect gene mutations with altered restriction sites, and can directly determine the genotype, but this method cannot be used for the detection of gene mutations without new restriction sites; while the traditional Solid-phase chips are expensive, and the sensitivity is not high, and the reproducibility of the test results is poor

Thirdly, both the detection method based on PCR technology (PCR-RFLP) and the allelic difference analysis method based on TaqMan technology have limitations in the detection throughput, and only one mutation can be detected at a time, which cannot meet the needs of practical applications.

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